Shrimp alkaline phosphatase

Abstract

The present invention relates to a nucleic acid molecule comprising a nucleotide sequence as set forth in SEQ ID Nos. 1 or 20 or having at least 55% sequence identify therewith and/or capable of hybridising under medium stringency conditions to the complementary sequence of SEQ ID Nos. 1 or 20, or the complementary sequence of said sequences, wherein said nucleotide sequence encodes or is complementary to a sequence which encodes a heat labile alkaline phosphatase and to a recombinant heat labile alkaline comprising: (a) all or a significant part of an amino acid sequence as shown in SEQ ID No. 2; or (b) all or a significant part of an amino acid sequence which has at least 60% sequence identify with SEQ ID No. 2 as well as methods for the manufacture thereof. 1 k AYWNK DAQDALDKQLGIKLREKQAKNVIFFLGDGMSLSTVTAARIYKGG  5ReRP6:17 51 LTGKFEREKISWEEFDFAALSKTYNTDKQ AYLTGVKTNQGV                               Active site 101 IGLDANTVRTNCSYQLDESLFTYSIAHWFQEAGRSTGVVTSTRVTHATPA 151 GTYAHVADRDWENDSDVVHDREDPEICDDIAEQLVFREPGKNFK VIMGGG                                         5ReRP6:26 201 RR GFFPEEALDIEDGIPGEREDGK HLITDWLDDK ASQGATASYVWNRDDL                          H23:30 251 LAVDIRNTDYLMGLFSYTHLDTVLTRDAEMDPTLPEMTKVAIEMLTKDEN 301 GFFLLVEGGRIDHMHHANQIRQSLAETLDMEEAVSMALSMTDPEETIILV 351 TADHGHTLTITGYADRNTDILDFAGISDLDDRRYTILDYGSGPGYHITED 401 GKRYEPTEEDLK DINFRYASAAPK HSVTHDGTDVGIWVNGPFAHLFTGVY               H23:18 451 EENYIPHALAYAACVGTGRTFCDEK *

1. An isolated nucleic acid molecule comprising a nucleotide sequence as set forth in any one of SEQ ID Nos. 1, 15, or 20 or having at least 95% sequence identity therewith, which encodes a heat labile alkaline phosphatase, or the complementary sequence of said sequences. 
     
     
       2. An isolated nucleic acid molecule comprising a nucleotide sequence which encodes a heat labile alkaline phosphatase, said alkaline phosphatase comprising the amino acid sequence of SEQ ID No. 2 or SEQ ID No. 16, or variants of said amino acid sequences which are at least 95% identical thereto or the complementary sequence of said nucleotide sequence. 
     
     
       3. The nucleic acid molecule of  claim 1  wherein the alkaline phosphatase is  Pandalus borealis  alkaline phosphatase. 
     
     
       4. A genetic construct comprising a nucleic acid molecule of  claim 1  operably linked to a promoter sequence. 
     
     
       5. A recombinant expression vector comprising a nucleic acid molecule of  claim 1  and which is capable of propagation in a host cell. 
     
     
       6. The vector of  claim 5  which is a plasmid or viral vector. 
     
     
       7. An isolated cell transformed with a construct or vector of  claim 4  or  5 . 
     
     
       8. The cell of  claim 7  which is a prokaryotic cell. 
     
     
       9. The cell of  claim 7  which is part of a eukaryotic cell culture. 
     
     
       10. An isolated recombinant cell or microorganism comprising a nucleic acid molecule of  claim 1 . 
     
     
       11. An isolated recombinant cell transformed with a nucleic acid molecule which encodes a heat labile alkaline phosphatase which comprises the amino acid sequence of SEQ ID No. 2 or SEQ ID No. 16 or an amino acid sequence which has at least 95% sequence identity with SEQ ID No. 2 or SEQ ID No. 16. 
     
     
       12. A method for the manufacture of heat labile alkaline phosphatase, comprising transforming a host cell with the vector of  claim 9 , maintaining the cell in a culture medium and isolating heat labile alkaline phosphatase expressed by said cell. 
     
     
       13. The nucleic acid molecule of  claim 1 , wherein said nucleic acid molecule comprises the nuecleotide sequence as set forth in SEQ ID No. 15. 
     
     
       14. The nucleic acid molecule of  claim 2 , wherein said alkaline phosphatase comprises the amino acid sequence as set forth in SEQ ID No. 16. 
     
     
       15. The nucleic acid molecule of  claim 1 , wherein said nucleic acid molecule comprises the nucleotide sequence as set forth in SEQ ID No. 1. 
     
     
       16. The nucleic acid molecule of  claim 1 , wherein said nucleic acid molecule comprises the nucleotide sequence as set forth in SEQ ID No. 20. 
     
     
       17. The nucleic acid molecule of  claim 2 , wherein said alkaline phosphatase comprises the amino acid sequence as set forth in SEQ ID No. 2.